Development of a High-Throughput Screen to Detect the Effects of Both Pre- and Post-Biotransformed Compounds for Enhanced Content Drug Discovery Workflows
Award last edited on: 12/21/2017

Sponsored Program
Awarding Agency
Total Award Amount
Award Phase
Solicitation Topic Code

Principal Investigator
Gary S Sayler

Company Information

490 Biotech Inc

2450 E J Chapman Drive
Knoxville, TN 37996
   (865) 974-9605
Location: Single
Congr. District: 02
County: Knox

Phase I

Contract Number: 1R43GM112241-01A1
Start Date: 7/1/2015    Completed: 12/31/2015
Phase I year
Phase I Amount
Development of a high-throughput screen to detect the effects of both pre- and post- biotransformed compounds for enhanced content drug discovery workflows Project Summary This Small Business Innovation Research Phase I project proposes to engineer a panel of autonomously bioluminescent human cell lines for the simultaneous, high-throughput detection of both pre- and post- biotransformed cytotoxicity onsets resulting from drug treatment across multiple tissue types to address the National Institute of General Medical Sciences (NIGMS) request for novel in vivo and in vitro methods for predicting the safety and toxicities of pharmacologic agents. With an average of 10,000 novel molecules that must be screened for each new lead compound developed, and an average of 10 to 15 years of research and development at a cost of up to $1B to manufacture one new drug, pharmaceutical companies must develop new testing regimens that provide more data at a lower cost in order to achieve the economics necessary to remain profitable. Up to 92% of failures for these new compounds at the clinical level are related to cytotoxicity, which often onl manifests during the costly and time consuming process of whole animal testing. This problem could be significantly mitigated by coordinately screening multiple tissue types simultaneously in a high-throughput fashion during upstream tier 1 screening. However, with existing bioluminescent reporter technology, this is simply not possible because the current market of bioluminescent reporter cells being applied toward toxicology screening relies upon a firefly luciferase gene construct that must be provided with a chemical substrate to activate its light emission response, resulting in only marginally informative single time point snapshots of potential toxicological interactions. In contrast, our substrate-free, autobioluminescent reporter cell lines emit light continuously and can modulate the output level of this signal in real time in response to cytotoxic interactions. This provides an uninterrupted stream of visual data over the lifetime of the reporter cell as it interacts and reacts to compound exposure at both its pre- and post-biotransformed states. Furthermore, by leveraging human cells as the sensing platform, our assay provides more accurate and realistic information in regards to bioavailability and a chemical's true effect on individual human health than does the employment of small animal models. With current in vitro screening assays now representing a $1.4B market with a predicted 12% annual growth rate, we believe we possess a product capable of significantly impacting the chemical/drug screening market and, here in particular, advancing our understanding of cytotoxic chemical biotransformations as they pertain to public health and consumer safety.

Public Health Relevance Statement:

Public Health Relevance:
The drug discovery process requires that upwards of 10,000 molecules be screened for each new lead compound developed and relies upon an expensive and time consuming combination of in vitro cell culture-based and in vivo whole animal-based models to identify, validate, and ensure the safety of any resulting potential therapeutic agents. This process is frustratingly exacerbated by an unfortunate dichotomy whereby the inexpensive in vitro cell culture systems used for tier 1 screening contribute to failures due to their inabiliy to model the complexity and parallel systems interaction inherent in whole animal models (which are responsible for up to 92% of new compound failures at the clinical level) and because they are not capable of demonstrating species-specific effects. To overcome these detractions and develop an improved tier 1 screening system that reduces the cost and time required for new compound evaluation, 490 BioTech proposes to develop a panel of multiple continuously bioluminescent human cell lines that will permit the simultaneous monitoring of each line to ascertain both the individual effects of compound treatment as well as the downstream effects of a compound's biotransformed metabolic breakdown products in real-time.

Project Terms:
Address; Animal Model; Animal Testing; Animals; Bacterial Luciferases; base; Biological Assay; Biological Availability; Biological Markers; Bioluminescence; Brain Stem; Cell Culture System; Cell Culture Techniques; Cell Line; cell type; Cells; Chemicals; Clinical; Complex; cost; cytotoxic; cytotoxicity; Data; design; Detection; Development; diacetyldichlorofluorescein; drug discovery; Drug Metabolic Detoxication; Economics; Eligibility Determination; Employment; Engineering; Ensure; Enzymes; Equipment; Evaluation; Failure (biologic function); Firefly Luciferases; Glutathione; Growth; Health; Heart; high throughput screening; Histocompatibility Testing; Human; Human Cell Line; human tissue; improved; In Vitro; in vivo; Individual; Lead; Light; light emission; Luc Gene; luciferin; Lung; Mandatory Testing; Marketing; Metabolic; Metabolic Biotransformation; Methods; Modeling; Monitor; National Institute of General Medical Sciences; novel; Organism; Output; Pancreas; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Positioning Attribute; Preclinical Drug Evaluation; Process; programs; public health medicine (field); public health relevance; Reaction; Reactive Oxygen Species; Regimen; Reporter; research and development; research clinical testing; response; Reverse Transcription; Safety; screening; Screening procedure; Series; Signal Transduction; Small Business Innovation Research Grant; Staging; Stem cells; Stream; System; T47D; Technology; Testing; Therapeutic; Therapeutic Agents; Time; Toxic effect; Toxicology; Transcript; transcriptomics; Variant; Visual

Phase II

Contract Number: 2R44GM112241-02
Start Date: 00/00/00    Completed: 00/00/00
Phase II year
(last award dollars: 2018)
Phase II Amount

Development of a high-throughput screen to detect the effects of both pre- and post- biotransformed compounds for enhanced content drug discovery workflows Project Summary This Small Business Innovation Research Phase II project will build upon our successful Phase I demonstration that substrate-free autobioluminescent signal generation can detect both the pre- and post-biotransformed metabolic impacts of therapeutic compounds from a single plate-based assay. Here, we will leverage this technology to develop a panel of industry-relevant autobioluminescent cell lines optimized for the detection of pre- and post-biotransformed compound metabolic impacts and the identification of specific detoxification pathway activation using modern three-dimensional (3D) microphysiological culture systems. These products and their underlying technology will specifically address the National Institute of General Medical Sciences (NIGMS) request for novel in vivo and in vitro methods for predicting the safety and toxicities of pharmacologic agents. By optimizing this technology to function within the industry-preferred 3D microphysiological format, we will address the critical need for new methods that can both identify compound toxicity and elucidate the mechanisms through which cells mitigate the compounds’ effects. The autonomous nature of this technology will increase toxicological data acquisition while preserving the critical advantage of presenting physiologically- relevant data, and reducing the cost of performance by eliminating substrates, reducing complexity, limiting hands-on operation time, obviating the need for sample destruction, and reducing the potential for measurement error. Through the validation of this technology at a scale relevant to tier 1 drug discovery screening and its comparative analysis against the existing gold-standard ATP content assay, this revolutionary approach is poised to have a significant and immediate impact towards reducing the estimated $8B/year in unnecessary expenditures made by pharmaceutical companies during their development of the 48% of new compounds that fail at the Phase I clinical trial stage due to misidentification of toxicological effects during tier 1 screening. This is possible because, as demonstrated in our Phase I work, the use of our autobioluminescent technology overcomes the high economic and logistical costs of existing, traditionally-bioluminescent cell’s requisite chemical substrate addition, which must co-occur with each generation of signal, and the intensive hands-on time necessitated to scale cultures due to their requisite sample destruction concurrent with imaging. Similarly, our autobioluminescent technology also obviates the hurdles presented by fluorescent cell’s susceptibility to autofluorescent signal inhibition and their tendency to remain active during downturns in cellular metabolism or even after cell death. The technology and products developed in this effort will therefore be capable of significantly improving the throughput and effectiveness of microphysiological systems-based tier 1 compound screening to improve the efficiency and economics of new compound development, and ultimately, consumer safety. This will allow them to thrive in a microphysiological system market that is predicted to maintain a compound annual growth rate of 70% to exceed $1.3B globally by 2022.

Public Health Relevance Statement:
Project Narrative Failure to identify the toxicity of pharmaceutical compounds such as Merck’s Vioxx, Bayer’s Baycol, and Wyeth’s FenPhen have caused consumers to be injured or killed by the drugs that were supposed to help them, and resulted in negative publicity and legal fees costing billions of dollars for the companies that produced them. While pharmaceutical companies have significantly enhanced their early stage toxicity screening regimens by transitioning to complex cell culture systems that can better identify new compounds’ toxicological effects on the human body to avoid repeating these mistakes, the use of these systems has increased the cost of new drug development to a level that is unsustainable for both consumers and the companies themselves due to a lack of technologies capable of economically identifying toxicity in these modern systems. In this Phase II R&D effort, 490 BioTech proposes to implement a novel toxicological screening technology based upon a synthetic luciferase genetic construct that links cellular health to the autonomous production of light in the visible spectrum to improve toxicity data acquisition by increasing the throughput and reducing the cost of modern pharmacological development assays while preserving their increased level of safety.

Project Terms:
Address; Affect; assay development; base; Biological Assay; Biotechnology; Cardiac; Cell Culture System; Cell Culture Techniques; Cell Death; Cell Line; Cell model; cell type; Cells; cerivastatin; Chemicals; Coculture Techniques; comparative; Complex; cost; CYP1A2 gene; CYP2B6 gene; CYP3A4 gene; cytotoxic; Data; data acquisition; Detection; Development; Dimensions; drug development; drug discovery; Drug Industry; Drug Metabolic Detoxication; drug metabolism; Early Diagnosis; Economics; Effectiveness; EPHX1 gene; Epithelial; established cell line; Evaluation; Expenditure; Failure; Fees; Generations; Genetic; Gold; Growth; Health; high throughput screening; Human; Human Biology; Human body; Image; improved; In Vitro; in vivo; Industrialization; Industry; injured; Kidney; Killings; Legal; Libraries; Light; Link; Liver; Logistics; Luciferases; Measurement; Metabolic; Metabolism; Methods; Modernization; NAT1 gene; National Institute of General Medical Sciences; Nature; novel; novel therapeutics; operation; Organ; Output; pancreatic cell line; Pathway interactions; Performance; Pharmaceutical Preparations; Pharmacologic Substance; Pharmacology; Phase; Phase I Clinical Trials; Physiological; Physiology; Predisposition; Prodrugs; Production; Regimen; Reporter; research and development; Research Personnel; Rofecoxib; Safety; Sampling; scaffold; screening; Signal Transduction; Small Business Innovation Research Grant; System; Technology; technology validation; Therapeutic; three dimensional cell culture; Time; Tissues; Toxic effect; Toxicity Tests; Toxicology; Vascular Endothelium; Work